CAREER: Uncovering Novel Factors for Multipass Membrane Protein Biogenesis

该奖项的全部或部分资金来自《2021年美国救援计划法案》(公法117-2)。这项工作的目标是了解人类细胞中一类生物必需的蛋白质--膜蛋白是如何形成的。所有的细胞都被一层薄薄的保护性“皮肤”包裹着，这种“皮肤”被称为膜，上面布满了蛋白质。这些膜蛋白允许细胞传递和接收信息和营养，因此对生物体如何对环境变化做出反应至关重要。因为这些膜蛋白含有蜡状的疏水性区域，必须被引导到这些膜上并嵌入其中，所以它们需要专门的机械来合成。这个项目试图了解用来制造这些膜蛋白的细胞机制。与这项研究相结合，将使用一项综合教育和推广计划，以突出被遗忘的妇女和代表不足的少数群体对科学的贡献，并增加不同学生接触生物研究的机会。美国救援计划的资金将支持这位研究人员在她职业生涯的关键阶段。膜蛋白的决定性结构特征是存在一个或多个跨膜结构域(TMD)，必须插入到脂质双层中。对于多通道膜蛋白，这些TMD在长度和疏水性上有很大的不同，因此它们的生物发生依赖于一组不完全了解的因素。该项目旨在确定和表征在人类细胞中插入、折叠和稳定不同的多通道膜蛋白所需的分子成员。新的膜蛋白生物发生因子将通过全基因组遗传筛选和对一组新生膜蛋白相互作用组的生化分析来确定。我们将结合功能和结构策略，利用体外重组和单粒子冷冻电子显微镜来剖析这些因子在膜蛋白生物发生中的分子作用。这项研究将建立膜蛋白生物发生的一般原则，这将适用于所有生命王国。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This award is funded in whole or in part under the American Rescue Plan Act of 2021 (Public Law 117-2).The goal of this work is to understand how a class of biologically essential proteins known as ‘membrane proteins’ are made in human cells. All cells are surrounded by a thin protective ‘skin’ known as a membrane, which is studded with proteins. These membrane proteins allow cells to both transmit and receive information and nutrients, and are thus are critical for how organisms respond and react to changes in their environment. Because these membrane proteins contain waxy, hydrophobic regions that must be guided-to and embedded within these membranes, they require specialized machinery for their synthesis. This project seeks to understand the cellular machinery used to make these membrane proteins. Coupled with this research, an integrated educational and outreach plan will be used to both highlight forgotten contributions from women and underrepresented minorities to science, and increase exposure of diverse students to biological research. American Rescue Plan funding will support this investigator at a critical stage in her career.The defining structural feature of a membrane protein is the presence of one or more transmembrane domains (TMDs) that must be inserted into the lipid bilayer. For multipass membrane proteins, these TMDs differ substantially in length and hydrophobicity, and thus rely on an incompletely understood set of factors for their biogenesis. This project seeks to identify and characterize the molecular players required for the insertion, folding, and stabilization of diverse multipass membrane proteins in human cells. Novel membrane protein biogenesis factors will be identified using genome wide genetic screens and biochemical analysis of the interactome of a panel of nascent membrane proteins. A combination of functional and structural strategies will be used to dissect the molecular role of these factors in membrane protein biogenesis using both in vitro reconstitution and single particle cryoelectron microscopy. This research will establish general principles of membrane protein biogenesis that will be relevant across all kingdoms of life.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.